Early Life Stages Research Articles

Climate challenges for fish larvae: Interactive multi-stressor effects impair acclimation potential of Atlantic herring larvae

Fish early life stages are particularly vulnerable and heavily affected by changing environmental factors. The interactive effects of multiple climate change-related stressors on fish larvae remain, however, largely underexplored. As rising temperatures can increase the abundance and virulence of bacteria, we investigated the combination of a spring heat wave and bacterial exposure on the development of Atlantic herring larvae (Clupea harengus). Eggs and larvae of Western Baltic Spring-spawners were reared at a normal and high temperature ramp and exposed to Vibrio alginolyticus and V. anguillarum, respectively. Subsequently, mRNA and miRNA transcriptomes, microbiota composition, growth and survival were assessed. Both high temperature and V. alginolyticus exposure induced a major downregulation of gene expression likely impeding larval cell proliferation. In contrast, interactive effects of elevated temperature and V. alginolyticus resulted in minimal gene expression changes, indicating an impaired plastic response, which may cause cellular damage reducing survival in later larval stages. The heat wave alone or in combination with V. alginolyticus induced a notable shift in miRNA expression leading to the down- but also upregulation of predicted target genes. Moreover, both increased temperature and the Vibrio exposures significantly altered the larval microbiota composition, with warming reducing microbial richness and diversity. The outcomes of this study highlight the high sensitivity of herring early life stages towards multiple climate change-related stressors. Our results indicate that interactive effects of rapidly changing environmental factors may exceed the larval stress threshold impairing essential acclimation responses, which may contribute to the ongoing recruitment decline of Western Baltic Spring-Spawning herring.

Additional treatments after curettage of congenital melanocytic nevi in the craniofacial region: A report from a single center in Japan

BackgroundCongenital melanocytic nevus (CMN) is a benign skin lesion present from birth, which may present with a risk of malignant transformation if extensive. Curettage, a treatment method involving the removal of the superficial layer of the nevus, is often used in the early stages of life. However, recurrence of the nevus and postoperative scarring may present as problems. Additional treatments, such as resection and/or laser treatment, are regularly required after curettage, particularly in the craniofacial region. However, no systematic treatment strategy has been reported. This study investigated additional treatments used after curettage to treat CMN in the craniofacial region and compared the frequency of treatments with respect to specific sites. MethodsCMN cases involving curettage as an initial treatment were retrospectively reviewed at Kyoto University Hospital between May 2019 and April 2022. ResultsThis study comprised 23 cases. Curettage was performed at a mean of 3.8 (1–10) months of age. No additional treatments were provided for 80% of head CMN. Additional treatments were performed in all cases, including the forehead and cheek. Laser treatment was performed in 86% of eyelid CMN and 75% of nasal CMN. Tissue expansion and flap closure were used in 33% of forehead CMN and 33% of cheek CMN. ConclusionsAdditional treatments used for CMN in the craniofacial region varied in accordance with the lesion site.

Exposure to thimerosal induces behavioral abnormality in the early life stages of zebrafish via altering amino acid homeostasis

Thimerosal (THI) has become a significant source of organic mercury pollutants in aquatic ecosystems, but there is limited information regarding its adverse effects on fish. In this study, zebrafish embryos were exposed to THI at 0 (control), 5.0, and 50 ng/L from 0–5 days post fertilization (dpf), and variations in their survival, development, behavior, free amino acid contents, and the biochemical responses involved in monoaminergic systems were examined. Although THI exposure did not significantly affect the survival, heart rate, or hatching time of zebrafish embryos, it substantially increased swimming velocity (136–154 % of the control) and reduced exploratory behavior (141–142 % of the control) in zebrafish larvae at 5 dpf. Exposure also significantly altered the amino acid contents (51–209 % of the control) and monoamine levels (70–154 % of the control) in zebrafish larvae, some of which displayed significant correlations with behavioral traits. THI significantly elevated dopamine receptor gene expression and monoamine oxidase activity in zebrafish larvae. Adding extra phenylalanine or tryptophan to the E3 medium facilitates the recovery of zebrafish larvae from the abnormal behaviors induced by THI. These findings reveal for the first time that THI exposure at the level of ng/L is sufficient to induce neurobehavioral toxic effects in the early life stages of zebrafish, and disrupting amino acid homeostasis is a critical underlying mechanism. This study provides valuable insights into the toxicity of THI to fish and highlights the importance of assessing its potential risks to aquatic ecosystems.

Dental replacement in Caipirasuchus (Crocodyliformes) from the Brazilian Cretaceous

Investigation on dental replacement patterns in three species of Caipirasuchus (Caipirasuchus paulistanus Iori and Carvalho, 2011; Caipirasuchus montealtensis (Andrade and bertini, 2008) and Caipirasuchus mineirus Martinelli et al., 2018), a Crocodyliformes from the Upper Cretaceous of Bauru Basin – Brazil, indicate aspects of their life habits. Caipirasuchus exhibits a posterior-to-anterior replacement teeth direction and numerical alternation in the teeth pattern. Remarkably, Caipirasuchus montealtensis shows replacements occurring even when the functional tooth is absent, possibly due to feeding or combat-related activities. Additionally, an intriguing inverted mandibular replacement in Caipirasuchus montealtensis suggests a unique teeth developmental process. On the other hand, Caipirasuchus mineirus exhibits minimal or no tooth replacements, suggesting a decreased need for dental maintenance in the early stages of life. Despite similarities in replacement patterns, further research is necessary to fully elucidate the lifestyle implications of these findings. This analysis underscores the importance of dental replacement studies in sphagesaurids for understanding their ecological adaptations.

Seasonal patterns of juvenile fish assemblages in the surf zones of tropical sandy beaches along Gaolong Bay, Hainan Island, China

AbstractAimSurf zones are crucial nursery habitats for the early life stages of fish species associated with typical coastal ecosystems. However, little is known about the temporal patterns and drivers of fish assemblages in tropical surf zones. This study aimed to assess the (1) main changes in fish community structure throughout 1 year, (2) seasonal dynamic patterns in fish assemblages, and (3) key factors influencing fish assemblages in the tropical surf zones.LocationGaolong Bay, Wenchang City, Hainan Island, China.MethodsFish sampling was conducted monthly from June 2021 to May 2022 using a beach seine net. Fish species were identified using both morphological and molecular analyses. Kruskal–Wallis test, analysis of similarity, non‐metric multidimensional scaling analysis, and similarity percentage analysis were used to investigate the temporal fish assemblage patterns. Generalised additive models and canonical correspondence analysis were used to assess how environmental variables influence fish assemblages.ResultsWe identified 83 fish species, which were grouped into three ecotypes based on their primary habitat: coral reef‐seagrass‐associated species (CS) (35), mangrove‐estuarine‐associated species (ME) (30), and common coastal‐estuarine‐associated species (CE) (18). Most captured individuals were juveniles, and fish abundance and diversity were highest in May. Most CS species were abundant between March and May. ME and most CE species were dominant from June to August, and Mugilidae (CE) was abundant between October and February. Furthermore, surf fish assemblages were substantially influenced by tidal level, water temperature, conductivity, pH, turbidity, and dissolved oxygen.ConclusionsJuvenile fish were abundant in May and fish species with three ecotypes alternate in the surf zones throughout the year. Counter to much current thinking, March maybe the spawn peak of most fish species in the studied area, and we suggest that the fishing ban period could start from March instead of May in the inshore areas of Hainan Island.

Evaluation of potential factors affecting the success of Cisco re-introduction and re-establishment in Lake Huron

Recent food web changes led to an opportunity to reintroduce (fingerling stocking) Cisco in central Lake Huron. We examined 9 factors affecting the ability of stocked Cisco fingerlings to survive, and reproduce and for their progeny to survive and reproduce. These values (obtained from the literature) were applied to a stochastic Leslie Matrix model and projected forward 30 years. The mean terminal eigenvalue (λ) of the final 10 years was used to indicate if the resulting wild population would continue to increase or decrease and thresholds for each variable. Recovery success was sensitive to survival rates especially in early life stages. Numbers stocked and the rate of homing by hatchery fish only affected the rate of recovery. Adult survival, if adjusted up, helped compensate for many of the earlier life stages lower survival rates. Simulations indicated the likelihood of successful establishment of ongoing natural reproduction if periodicity of boom years of reproduction were at least once every 5 years on average, but higher adult survival rates can relax this requirement allowing for an expanding population at less frequent periodicities.

Comparative Study of the Effects of Drugs Targeting Adrenergic Receptors on the Early Life Stages of Zebrafish.

Owing to the presence of drugs targeting adrenergic receptors in aquatic ecosystems, considerable attention has been directed towards their environmental distribution and fate in recent decades. However, their potential impacts on non-target aquatic organisms, particularly fish, have received relatively limited investigation. In this study, moxisylyte (MOX) and propranolol (PRO) were selected as representatives of α- or β-adrenergic receptor antagonist, respectively, and we assessed their effects on the early life stages of zebrafish, especially on the nervous and cardiovascular systems. Although both compounds exhibited marginal effects on zebrafish survival, hatching and gross abnormality following exposure to concentrations ranging from 1 to 625 μg/L, they adversely affected the development of cardiovascular and nervous systems, but through different mechanisms of action, as evidenced by variations in gene transcriptional responses and enzyme activities. Notably, cardiovascular responses appear promising for use as potential biomarkers for exposure to drugs targeting adrenergic receptors. This study enhances our understanding of the ecotoxicological risks posed by α- and β-blockers in fish. Nonetheless, further investigation is needed to elucidate the precise mechanisms underlying the impacts of drugs targeting adrenergic receptors due to our limited knowledge of the physiological functions of the adrenergic system in fish.

Life stage and vaccination shape the gut microbiome of hatchery-reared Atlantic salmon (Salmo salar) intended for river stocking

Microbiomes play an essential role in promoting host health and fitness in fishes, but the factors driving variation in gut microbiomes among individuals are not fully recognized. Understanding the biological and environmental factors influencing variation in the gut microbiome in fishes is needed to link microbiome composition to host physiology and potentially design manipulations to improve rearing success in fisheries and aquaculture. In this study, we characterized the gut microbiomes of Atlantic salmon (Salmo salar), a globally important species in commercial aquaculture and recreational fishing, considering life stage, vaccination status, and hatchery origin. Microbiomes diverged between hatchery sub-adults (age-0 parrs) and wild adults, with sub-adults exhibiting higher diversity and more similar microbiome communities when compared to adults. The diversity and composition of gut microbiomes did not differ between groups of adults that were reared in two different hatcheries and sampled from the wild. We also found that oral vaccines against bacterial kidney disease (BKD) reduced gut microbial diversity within individual sub-adult salmon, driving increased differentiation of gut microbial communities among individuals. Unvaccinated sub-adults displayed a stable core gut microbiome that was greatly reduced upon vaccination for BKD. Our results suggest that life stage is an essential factor shaping the gut microbiome of Atlantic salmon, likely driven by differences in habitat and diet, and that BKD vaccination can lead to a rapid disruption of the gut microbiome. Conditions experienced in the hatchery environment appear to have a strong influence on host gut microbiomes, but differences at early life stages may be lost due to environmental factors experienced in the wild. The plasticity of the gut microbiome throughout the life of individuals may have important implications for understanding host health, with potential applications for improving the rearing and reintroduction success of the ecologically and economically important Atlantic salmon.

Understanding the dynamics of fish spawning phenology and habitat in a changing ecosystem using a long-term ichthyoplankton monitoring dataset

As ecosystems change, understanding the consequences for fish population dynamics and habitat use is essential for resource management. Using white perch ( Morone americana) survey data on early life stages collected during a long-term ichthyoplankton monitoring program in the Hudson River (New York, USA), an ecosystem under immense pressure from climate change, ecological shifts, and anthropogenic activities, we evaluated the drivers of changes in egg abundance and spawning habitat between 1980 and 2017. Results indicated that egg abundance is associated nonlinearly with temperature, conductivity, discharge, depth, location, and the week of year. We also found that egg abundance has declined within the river over time. Additionally, shifts in hotspots of spawning activity were identified, including evidence that the lower extent of spawning habitat has moved upriver since 1980. This study indicates that the dynamics of white perch early life histories and spawning habitat are changing. It also highlights the utility of a long-term monitoring program for broadening our understanding of fish ecology in the age of big data in changing ecosystems.

Nanoplastics enhanced the developmental toxicity of aromatic disinfection byproducts to a marine polychaete at non-feeding early life stage

Micro/nanoplastics can act as vectors for organic pollutants and enhance their toxicity, which has been attributed to the ingestion by organisms and the “Trojan horse effect”. In this study, we disclosed a non-ingestion pathway for the toxicity enhancement effect of nanoplastics. Initially, the combined toxicity of polystyrene microplastics (40 μm) or nanoplastics (50 nm) with three disinfection byproducts (DBPs) to a marine polychaete, Platynereis dumerilii, was investigated. No toxic effect was observed for the micro/nanoplastics alone. The microplastics showed no effect on the toxicity of the three DBPs, whereas the nanoplastics significantly enhanced the toxicity of two aromatic DBPs when the polychaete was in its non-feeding early life stage throughout the exposure period. The microplastics showed no interaction with the P. dumerilii embryos, whereas the nanoplastics agglomerated strongly on the embryonic chorion and fully encapsulated the embryos. This could contribute to higher actual exposure concentrations in the microenvironment around the embryos, as the concentrations of the two aromatic DBPs on the nanoplastics were 1200 and 120 times higher than those in bulk solution. Our findings highlight an important and previously overlooked mechanism by which nanoplastics and organic pollutants, such as DBPs, pose a higher risk to marine species at their vulnerable early life stages. This study may contribute to a broader understanding of the environmental impacts of plastic pollution and underscore the necessity to mitigate their risks associated with DBPs.

Developmental plasticity of the cardiovascular system in oviparous vertebrates: effects of chronic hypoxia and interactive stressors in the context of climate change.

Animals at early life stages are generally more sensitive to environmental stress than adults. This is especially true of oviparous vertebrates that develop in variable environments with little or no parental care. These organisms regularly experience environmental fluctuations as part of their natural development, but climate change is increasing the frequency and intensity of these events. The developmental plasticity of oviparous vertebrates will therefore play a critical role in determining their future fitness and survival. In this Review, we discuss and compare the phenotypic consequences of chronic developmental hypoxia on the cardiovascular system of oviparous vertebrates. In particular, we focus on species-specific responses, critical windows, thresholds for responses and the interactive effects of other stressors, such as temperature and hypercapnia. Although important progress has been made, our Review identifies knowledge gaps that need to be addressed if we are to fully understand the impact of climate change on the developmental plasticity of the oviparous vertebrate cardiovascular system.

Integrated Multi-Biomarker Responses of Juvenile Zebra Seabream (Diplodus cervinus) to Warming and Acidification Conditions

The impacts of climate change-related stressors are becoming more noticeable in the ocean, particularly in coastal marine ecosystems. Yet limited information still exists on the physiological state and ecological resilience of marine fish species, especially during their early life stages (i.e., larvae and juveniles). The present study investigated the effects of chronic exposure to seawater warming (OW; ΔT = +4 °C) and acidification (OA; ΔpH = −0.3 pH units, equivalent to pCO2~1000 µatm), acting alone or combined (OWA), on juvenile zebra seabream (Diplodus cervinus) physiological resilience, considering distinct levels of biological organization (i.e., biochemical, cell, organ and individual levels). After 60 days of exposure, both stressors, in isolation or combination, significantly decreased specific growth rate (−11% in OW, −42% in OA and −49% in OWA) and leukocyte counts (from −29% in OA and OWA up to −37% in OW) in relation to the control treatment. In addition, a decreased Fulton’s condition index (K) was observed under warming and acidification in combination (−35% in OWA). At the cell level, OW, OA and OWA triggered different biomarker responses in D. cervinus (i.e., up-regulation, down-regulation, or absence of significant effect). In general, the results are suggestive of an antagonistic effect when warming and acidification are combined. OWA yielded the highest integrated biomarker response (IBR) index value in the whole organism, muscle, brain and gills of D. cervinus juveniles, therefore suggesting that the effects of these stressors are more severe when they act together. The distinct patterns observed in each stress scenario highlight the importance of carrying out further studies adjusted to the specificities of different regions, i.e., accounting not only for the type and degree of severity of environmental stressors already felt and/or projected for that specific area, but also the physiological plasticity of species that inhabit a particular ecosystem. The gathered knowledge will allow one to determine the vulnerability of particular marine species and geographic areas and, most importantly, to draw up effective and tailor-made conservation strategies to overcome climate change impacts.

Water-accommodated fractions of heavy and light oils impact DNA integrity, embryonic development, and immune system of Japanese medaka at early life stages.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants generally found in complex mixtures. PAHs are known to cause pleiotropic effects on living organisms, including developmental defects, mutagenicity, carcinogenicity and immunotoxicity, and endocrine disruptions. The main goal of this study is to evaluate the toxicity of water-accommodated fractions (WAFs) of oils in two life stages of the Japanese medaka, larvae and juveniles. The deleterious effects of an acute exposure of 48h to two WAFs from Arabian light crude oil (LO) and refined oil from Erika (HO) were analyzed in both stages. Relevant endpoints, including ethoxy resorufin-O-deethylase (EROD) activity, DNA damage (Comet assay), photomotor response, and sensitivity to nervous necrosis virus (NNV) infection, were investigated. Larvae exposed to both oil WAFs displayed a significant induction of EROD activity, DNA damage, and developmental anomalies, but no behavioral changes. Deleterious effects were significantly increased following exposure to 1 and 10μg/L of LO WAFs and 10μg/L of HO WAFs. Larval infection to NNV induced fish mortality and sharply reduced reaction to light stimulation. Co-exposure to WAFs and NNV increased the mortality rate, suggesting an impact of WAFs on fish defense capacities. WAF toxicity on juveniles was only observed following the NNV challenge, with a higher sensitivity to HO WAFs than to LO WAFs. This study highlighted that environmentally realistic exposure to oil WAFs containing different compositions and concentrations of oil generated high adverse effects, especially in the larval stage. This kind of multi-marker approach is particularly relevant to characterize the toxicity fingerprint of environmental mixtures of hydrocarbons and PAHs.

Novel high-throughput oxygen saturation measurements for quantifying the physiological performance of macroalgal early life stages.

Understanding how macroalgal forests will respond to environmental change is critical for predicting future impacts on coastal ecosystems. Although measures of adult macroalgae physiological responses to environmental stress are advancing, measures of early life-stage physiology are rare, in part due to the methodological difficulties associated with their small size. Here we tested a novel, high-throughput method (rate of oxygen consumption and production; ) via a sensor dish reader microplate system to rapidly measure physiological rates of the early life stages of three habitat-forming macroalgae, the kelp Ecklonia radiata and the fucoids Hormosira banksii and Phyllospora comosa. We measured the rate of O2 consumption (respiration) and O2 production (net primary production) to then calculate gross primary production (GPP) under temperatures representing their natural thermal range. The microplate system was suitable for rapidly measuring physiological rates over a temperature gradient to establish thermal performance curves for all species. The microplate system proved efficient for measures of early life stages of macroalgae ranging in size from approximately 50 μm up to 150 mm. This method has the potential for measuring responses of early life stages across a range of environmental factors, species, populations, and developmental stages, vastly increasing the speed, precision, and efficacy of macroalgal physiological measures under future ocean change scenarios.

Thermal performance of native and invasive crab species: investigating the invasion potential of Hemigrapsus takanoi in southern European Carcinus maenas’ habitats

Invasive species’ plasticity and latitudinal spread have become an increasing matter of concern in rapidly changing climate. The brush-clawed shore crab (Hemigrapsus takanoi), native along northwestern Pacific shores, has established dense populations in northern European coasts. Detrimental effects in natura have been reported, such as displacement of native species and competition for shelter, rasing concerns regarding shellfish early life stages and food webs-posing a threat to wild stocks’ recruitment and aquaculture. The species has not yet been observed in southern European countries (e.g., Portugal), where niche habitats such as harbours, mudflats, and lagoons reach higher temperatures than the invaded northern range. The aims of this study were to determine H. takanoi potential to acclimate to a southern thermal range, assess thermal performance, and identify potential competitive advantages over the native shore crab Carcinus maenas. This was accomplished through individual exposure to one of three treatments, resembling a southern European thermal range (17 °C, 21 °C, or 25 °C), for 30 days. H. takanoi thrived in temperatures as high as 25 °C, a realistic thermal scenario in Óbidos Lagoon (the biggest Portuguese lagoon)—while the native shore crab C. maenas underperformed at that treatment, showing lower survival, growth, and feed intake than H. takanoi. These results suggest that H. takanoi may be able to colonise southern and warmer European locations and have higher plasticity than the native species already inhabiting areas close to its upper thermal limits. Under global warming scenarios, inferences of further latitudinal spread and increased invaded range may also be taken to the invaded northern European habitats.

Multiple climate change stressors reduce the emergence success of gravel-spawning fish species and alter temporal emergence patterns

Climate change, with its profound effects on stream sediment, hydrological, and temperature dynamics, will exacerbate impacts on habitat conditions for many species, particularly those with vulnerable early life stages relying on the hyporheic zone, such as gravel-spawning fishes. Due to the complex and interactive nature of multiple stressor effects, we employed large-scale outdoor mesocosms to systemically test how the reproductive success of three gravel-spawning fish species brown trout (Salmo trutta), nase, (Chrondrostoma nasus) and Danube salmon (Hucho hucho) was affected by individual and combined effects of warming (+3–4 °C), fine sediment (increase in <0.85 mm by 22 %) and low-flow (eightfold discharge-reduction). Fine sediment had the most detrimental effect on emergence rate and fry length in all three species, reducing the emergence rate to zero in brown trout, 9 % in nase, and 4 % in Danube salmon. The emergence mortality caused by fine sediment surpassed that of hatching distinctly, suggesting that negative effects due to hypoxia were considerably exacerbated by entombment. Warming had only minor effects as a single stressor, but low flow reduced emergence rates of the spring spawning species nase and Danube salmon by 8 and 50 %, respectively. In combined treatments including fine sediment, however, the emergence success of all three species responded strongly negatively, even in the cyprinid species nase, which showed little interactive effects between stressors regarding hatching success. Warming and fine sediment also led to the earlier emergence of fry, implying a risk of asynchrony with available food resources. This study dramatically shows that climate change can have deleterious impacts on the reproductive success of gravel-spawning fish species, irrespective of taxonomic or ecological traits.

Hyper IgE Syndromes.

The Hyper IgE Syndromes are rare primary immunodeficiencies characterized by eczema, recurrent skin and respiratory infections and elevated serum IgE levels. Nowadays a geneticmolecular characterization is possible and allows the distinction in various monogenic pathologies, which share some clinical characteristics but also important differences. In addition to long-known STAT3 and DOCK8 gene mutations, in fact, also ZNF341, CARD11, ERBB2IP, IL6R and IL6ST genes mutations can cause the disease. The main clinical manifestations are represented by newborn rash, eczema similar to atopic dermatitis, bacterial and viral skin infections, cold abscesses, respiratory infections with possible pulmonary complications, allergies, gastrointestinal manifestations, malignancies and connective tissue abnormalities. Diagnosis is still a challenge because, especially in the early stages of life, it is difficult to distinguish from other pathologies characterized by eczema and high IgE, such as atopic dermatitis. Several scores and diagnostic pathways have been developed, but it is essential to seek a genetic diagnosis. Treatment is based on prevention and early treatment of infections, meticulous skincare, intravenous immunoglobulins and HSCT, which, in some HIES subtypes, can modify the prognosis. Prognosis is related to the affected gene, but also to early diagnosis, timely treatment of infections and early HSCT.

Early-life diet composition affects phenotypic variation of correlated animal personality traits.

Behavioural traits are under both genetic and environmental influence during early life stages. Early environmental conditions related to the amount and type of food have been found to alter behaviour in many organisms. However, how early life diet affects the variation in and the correlation between behavioural traits is largely unknown. Using a multivariate approach, we investigated how variation in parental prey selection is related to three repeatable nestling personality traits, and explored the within and between-individual covariation between these behaviours in a wild passerine, the great tit (Parus major). Our results confirm that breath rate, docility and handling aggression (HA) in great tit nestlings are repeatable traits. Contrary to our expectation, the three nestling personality traits did not form a behavioural 'syndrome' on the phenotypic level in the study population, but we found two of three expected phenotypic correlations, mostly at the within-individual level. Moreover, we found that breath rate significantly decreased with a higher number of spiders in the diet, and docility and handling aggression were significantly and inversely related to higher numbers of noctuids and tortricids in the diets of individuals within broods. Thus, our findings suggest that provisioning quantity and quality during the early life, affects variation in behavioural phenotypes, which occurs mainly at the within-individual level.

Effects of climate change on seed germination may contribute to habitat homogenization in freshwater forested wetlands

Climate changes in temperate regions are expected to result in warmer, shorter winters in temperate latitudes. These changes may have consequences for germination of plant species that require a period of physiological dormancy. The effect of cold duration on seed germination has been investigated in a number of plant taxa, but has not been well studied in wetland and bottomland forest tree species, an ecosystem that is threatened by habitat homogenization. Our work sought to test the role of changing winter temperatures on seed germination in specialist (Nyssa aquatica and Taxodium distichum) and generalist (Acer rubrum and Liquidambar styraciflua) tree species within forested wetlands throughout the eastern U.S. The experiment was conducted in an environmental chamber in Norfolk, VA, USA. Seeds of T. distichum, N. aquatica, A. rubrum, and L. styraciflua were exposed to each of seven pre-germination cold exposure durations (0, 15, 30, 45, 60, 75, and 90 days) and observed for germination for 30 days. Cold stratification duration positively impacted total percent germination in N. aquatica (p < 0.0001) as well as A. rubrum (p = 0.0008) and T. distichum (p = 0.05). Liquidambar styraciflua seeds exhibited more rapid rates of germination with increasing cold exposure duration and greater percent germination compared to the others regardless of cold stratification duration. Our results provide insight into how community dynamics and biodiversity of wetland and bottomland trees may shift with a changing climate. Further, this work emphasizes the importance of understanding the role of plant functional traits in early life stages in community dynamics and has implications for management practices.

Cross-sectional and longitudinal evaluation of heart-to-brachium pulse wave velocity for cardiovascular disease risk.

Heart-brachium pulse wave velocity (hbPWV) is a promising measure of arterial stiffness including the proximal aorta. To characterize age-associated changes and the clinical utilities of hbPWV, we evaluated the impacts of age and cardiovascular disease (CVD) risks on hbPWV cross-sectionally (N = 7868) and longitudinally (N = 3710, followed by 9.1 ± 2.0 years). hbPWV were obtained using two validated equations for arterial path length (with and without considering age-related aortic elongations). Brachial-ankle pulse wave velocity (baPWV) was used as a comparative measure. Repeated-measures correlation (rmcorr) and regression analyses were used to characterize associations of PWVs with age and Framingham's general CVD risk score (FRS). In the cross-sectional study, hbPWVs derived by both equations showed stronger correlation with age (r = 0.746 ~ 0.796) and FRS (r = 0.714-0.749) than baPWV (r = 0.554 and r = 0.643). Furthermore, hbPWVs correlated with FRS even after controlling for age (r = 0.260 ~ 0.269, P < 0.0001). In the longitudinal study, hbPWVs demonstrated significantly higher rmcorr coefficient with age than baPWV (rrm=0.439-0.511 vs. 0.307, P < 0.0001). Across the adult lifespan, age-related increases in hbPWVs were almost consistent, starting from young adults, while baPWV displayed accelerated increases with age. A receiver operating characteristic curve analysis indicated that hbPWVs depicted more robust ability to stratify general CVD risk compared with baPWV (AUC = 0.896-0.913 vs. 0.833, P < 0.0001). The results of the follow-up study were consistent with the findings of the cross-sectional investigation. Our findings suggest that hbPWV undergoes a linear augmentation with age, commencing from an early adult life stage onward, rendering it a potential marker for discerning CVD risk.